Retinal dystrophies and variants in PRPH2 / Distrofias retinianas e variantes em PRPH2

ABSTRACT - This report presents three patients diagnosed with macular dystrophies with variants in PRPH2. Peripherin-2, the protein of this gene, is important in the morphogenesis and stabilization of the photoreceptor outer segment. Peripherin-2 deficiencies cause cellular apoptosis. Moreover, pathogenic variants in PRPH2 are associated with various diseases, such as pattern, butterfly-shaped pattern, central areolar, adult-onset vitelliform macular, and cone-rod dystrophies as well as retinitis pigmentosa, retinitis punctata albescens, Leber congenital amaurosis, fundus flavimaculatus, and Stargardt disease.

RESUMO - Este relato apresenta três pacientes com diagnóstico de distrofias maculares com mutações no PRPH2. Periferina 2, a proteína deste gene, é importante na morfogênese e estabilização do segmento externo dos fotorreceptores. Deficiências de periferina 2 causam apoptose celular. Além disso, variantes patogênicas no PRPH2 estão relacionadas a diferentes doenças, como distrofia padrão, distrofia padrão em asa de borboleta, distrofia central areolar, distrofia viteliforme do adulto, retinose pigmentar, distrofia de cones e bastonetes, retinite punctata albscens, amaurose congênita de Leber, fundus flavimaculatus e doença de Stargardt.

Vasculopathy associated with peripheral neuropathy in gait parameters of diabetic people

Abstract Diabetic peripheral neuropathy (DPN) is a common complication of diabetes mellitus when glycemic levels are poorly controlled. Sometimes DPN is accompanied by vasculopathy (DPV), which can worsen the clinical prognosis. The aim of this study was to analyze the gait parameters of nondiabetic individuals and diabetic individuals with DPN with or without DPV. METHOD The study included 68 individuals (50 to 65 years old) divided into three groups: people without diabetes mellitus (n = 33), diabetic patients with DPN (n = 18), and diabetic patients with both DPN and DVP (n = 17). The participants underwent a gait evaluation using electronic baropodometry to obtain the single and double support, velocity, and pressure-time integral. RESULTS The pressure-time integral, velocity, and single support variables were lower, and the double support and double support/single support ratio were higher in the diabetic neuropathy and vasculopathy group. The velocity was lower the greater the degree of impairment of the diabetic foot. Some correlations were identified with velocity. CONCLUSION In diabetic individuals, there was a significant worsening of the gait parameters analyzed according to increasing degree of clinical impairment.

Comparação dos efeitos da ventilação mecânica não invasiva contínua e intermitente sobre parâmetros cardiorrespiratórios e modulação autonômica de indivíduos saudáveis / Comparison of the effects of continuous and intermittent non invasive mechanical ventilation on cardiorespiratory parameters and autonomic modulation of healthy individuals

Modelo do estudo: Estudo transversal. Objetivo: Avaliar a influência da Ventilação Mecânica não Invasiva (VMNI) de forma contínua e intermitente sobre a modulação autonômica cardíaca e parâmetros cardiorrespiratórios em mulheres saudáveis. Métodos: Vinte voluntárias realizaram duas modalidades de VMNI: contínua por meio do CPAP e intermitente por meio do Reanimador de Muller. Inicialmente permaneceram em repouso em respiração espontânea por 20 minutos. Em seguida foram submetidas a 20 minutos de aplicação de VMNI com a técnica selecionada e por fim permaneceram 20 minutos em repouso em respiração espontânea. Os parâmetros cardiorrespiratórios e variabilidade da frequência cardíaca (VFC) foram mensurados em todos estes momentos. Resultados: Não houve diferenças significantes quando comparadas as duas técnicas. Observaram-se diferenças nos parâmetros cardiorrespiratórios e VFC quando analisadas individualmente. Menores valores de frequência cardíaca e frequência respiratória foram observados na modalidade contínua quando comparado os valores durante a ventilação com respiração espontânea (p<0,005). Em ambas técnicas observaram-se aumentos significantes de SpO2 durante a ventilação em comparação a respiração espontânea. Observou-se aumento da modulação parassimpática (RMSSD, HF ms2 e SD1) e da variabilidade global (SDNN, RR triangular e SD2) em ambas as técnicas quando comparado ventilação e respiração espontânea (p<0,005). Conclusão: Não houve diferença quando comparadas as duas modalidades de VMNI. Contudo, quando analisadas individualmente observam-se comportamento diferentes dos parâmetros cardiorrespiratórios e na modulação autonômica.

Study design: Cross-sectional study. Objective: To evaluate the influence of continuous and intermittent non invasive mechanical ventilation (NIV) on cardiac autonomic modulation and cardiorespiratory parameters in healthy women. Methods: Twenty subjects performed two types of NIV: continuous through CPAP and intermittent through Müller Reanimator. Initially they remained at rest for 20 minutes in spontaneous breathing. Then volunteers were subjected to 20 minutes of NIV application with the selected technique and finally remained at rest for 20 minutes in spontaneous breathing. Cardiorespiratory parameters and heart rate variability (HRV) were measured in all these moments. Results: Therewere no significant differences when comparing the two techniques. Differences were observed in HRVand cardiorespiratory parameters when analyzed individually. Lower values of heart rate and respiratory rate were observed in continuous ventilation when compared to values during ventilation with spontaneous breathing (p <0.005). In both techniques we observed significant increases in SpO2 during ventilation compared to spontaneous breathing. We observed an increase in parasympathetic modulation (RMSSD, HF ms2 and SD1) and overall variability (SDNN, RR triangular and SD2) in both techniques when compared ventilation to spontaneous breathing (p <0.005). Conclusion: There was no difference comparing the two types of NIV. However, when analyzed individually we observe different behavior of cardiorespiratory parameters and autonomic modulation.

Implementation of physiotherapeutic shares in the prevention of diabetes complications in a Family Health Strategy / Implementação de ações fisioterapêuticas na prevenção de complicações do diabetes na Estratégia de Saúde da Família

Introduction In the Family Health Strategy (FHS), the treatment of Diabetes Mellitus (DM) includes education and lifestyle change strategies. Physiotherapists have a key role in this health setting. Objectives To implement actions of evaluation and guidelines for patients with type 2 DM who attend a Family Health Strategy (FHS), regarding diabetic foot and the practice of regular physical exercise in the control and prevention of the complications of Diabetes Mellitus. Methods 17 individuals from an FHS were evaluated, with the following procedures: clinical and anthropometric parameters, inspection, a questionnaire on diabetic neuropathy, tests of vibratory and tactile sensitivity, muscle function, range of motion, functional analysis, questions about exercise practice and guidance regarding controlling blood glucose and foot care. Results Deformities, dry skin, calluses, dehydration, ulceration, cracking and brittle nails were found. Peripheral neuropathy was not observed; tactile sensitivity was altered in the heel region and the vibratory sense was absent in 5% of individuals. A decrease in functionality of ankle movements was verified. Of the participants, 76% were sedentary, 24% knew about the benefits of practicing regular exercise, 25% had undergone a medical evaluation prior to performing physical exercise and, of these, 25% were supervised by a qualified professional. Discussion The implementation of physiotherapy actions in diabetics from an FHS was important for highlighting the presence of risk factors for diabetic complications. Conclusions Individuals attending the FHS need more information and programs for the prevention of diabetic complications. .

Introdução Na Estratégia da Saúde da Família (ESF) o tratamento do Diabetes Mellitus (DM) inclui estratégias de educação e modificação do estilo de vida. Os fisioterapeutas vêm desenvolvendo papel fundamental nesse novo cenário de saúde. Objetivos Implementar ações de avaliação e orientações para pacientes com DM tipo 2, quanto ao pé diabético e à prática regular de exercícios físicos no controle e na prevenção de complicações do DM, os quais frequentam uma Estratégia de Saúde da Família (ESF). Métodos Foram avaliados 17 indivíduos de uma ESF, com os seguintes procedimentos: verificação de parâmetros clínicos e antropométricos, inspeção, questionário sobre neuropatia diabética, teste da sensibilidade tátil e vibratória, função muscular, amplitude de movimento, análise funcional, questionamentos sobre realização de exercícios, orientações sobre sua prática no controle glicêmico e orientações quanto aos cuidados com os pés. Resultados Foram encontradas deformidades, pele ressecada, calos, desidratação, ulceração, rachaduras e unhas quebradiças. A investigação sobre neuropatia periférica não indicou sua presença; a sensibilidade tátil estava alterada principalmente na região do calcanhar e a sensibilidade vibratória estava ausente em 5% dos indivíduos. Verificou-se diminuição na funcionalidade em movimentos de tornozelo. Dos participantes, 76% eram sedentários e 24% conheciam os benefícios da prática regular de exercícios físicos; 25% dos indivíduos passaram por avaliação médica antes de realizar exercício físico, que era supervisionado por profissional habilitado em 25% dos casos. Discussão A implementação de ações fisioterapêuticas nos diabéticos de uma EFS foi importante para destacar a presença de fatores de risco para complicações do diabetes. Conclusões Os indivíduos da ESF necessitam de mais informações e programas para a prevenção de complicações do DM e apresentam alterações predisponentes a amputações e outras complicações. .

Avaliação do fator CIITA como potencial adjuvante molecular para vacinas e imunoterapias / Evaluation of CIITA factor as a potential molecular adjuvant for vaccines and immunotherapies

O fator CIITA é a proteína responsável por controlar a transcrição de genes do complexo principal de histocompatibilidade de classe II (MHC II) envolvidos na apresentação antigênica a linfócitos T CD4+. A expressão desta proteína é complexa e célula-específica, dependendo de mecanismos de regulação transcricionais e póstranscricionais. Com o intuito de investigar o potencial do fator CIITA como adjuvante molecular, no presente estudo desenvolvemos e validamos sistemas de transferência gênica capazes de promover a eficiente expressão de CIITA em vários tipos celulares. Além disso, investigamos a regulação pós-traducional deste fator em células não hematopoéticas. Desta forma, foram produzidos um vetor plasmidial e um vetor lentiviral, ambos carreando a sequência do fator CIITA humano desenhada in silico visando a eliminação de elementos cis-reguladores, e otimizada para eficiente expressão em células humanas. A transfecção/transdução de três linhagens de células humanas não hematopoéticas resultou na eficiente expressão de CIITA com localização nuclear apropriada. Células expressando CIITA apresentaram síntese de novo do MHC II, confirmando a funcionalidade da proteína e validando ambos os vetores para a análise futura da atividade adjuvante do CIITA em imunizações gênicas. Ensaios preliminares de inoculação de explantes de pele humana com o vetor lentiviral evidenciaram a eficiente transdução e expressão do CIITA exógeno em células primárias. Em seguida, células dendríticas (DCs) derivadas de monócitos de indivíduos saudáveis ou infectados com HIV-1 foram transduzidas com o vetor lentiviral para confirmar a expressão do CIITA em células primárias e avaliar a aplicação desse sistema adjuvante no aprimoramento da vacina de DCs anti-HIV. DCs de indivíduos saudáveis ou infectados foram transduzidas com sucesso pelo lentivírus, o qual induziu uma produção prolongada do mRNA codificando CIITA. Entretanto, os vetores lentivirais induziram um...

The CIITA factor is a protein responsible for controlling the transcription of major histocompatibility complex class II (MHC II) genes involved on antigen presentation to CD4+ T helper cells. The expression of this transcription factor is complex and differs in various cell types depending on transcriptional and post-transcriptional regulatory mechanisms. In order to investigate the CIITA factor potential as molecular adjuvant, here we developed and validated two gene delivery systems capable of promoting efficient CIITA expression in various human cell types. Additionally, we applied the delivery systems to investigate the post-translational regulation of this factor in nonimmune cells. A DNA plasmid and a lentiviral vector were produced, both carrying the human CIITA DNA sequence in silico designed to avoid cis-regulatory elements, and genetic optimized for expression efficacy in human cells. Transfection or transduction of three different non-immune human cell lines resulted in efficient CIITA expression with proper nuclear localization. The CIITA-expressing cells presented de novo MHC II molecules expression confirming the functionality of the exogenous protein, and validating both delivery systems for the future analysis of the CIITA adjuvant activity in genetic immunizations. Preliminary assays involving the inoculation of the lentiviral vector into human skin explants showed efficient transduction and expression of exogenous CIITA in primary cells. Next, monocyte-derived dendritic cells (DCs) from healthy individuals and HIV-1-infected patients were transduced with the lentiviral vector to confirm the exogenous CIITA expression in primary human cells and also evaluate the applicability of this adjuvant system to improve the DC-based vaccines against HIV. DCs from healthy and infected individuals were successfully transduced by the lentivirus, which induced a sustained CIITA mRNA production. However, the vector particles by themselves induced an...

Membrane and envelope virus proteins co-expressed as lysosome associated membrane protein (LAMP) fused antigens: a potential tool to develop DNA vaccines against flaviviruses

Vaccination is the most practical and cost-effective strategy to prevent the majority of the flavivirus infection to which there is an available vaccine. However, vaccines based on attenuated virus can potentially promote collateral side effects and even rare fatal reactions. Given this scenario, the developent of alternative vaccination strategies such as DNA-based vaccines encoding specific flavivirus sequences are being considered. Endogenous cytoplasmic antigens, characteristically plasmid DNA-vaccine encoded, are mainly presented to the immune system through Major Histocompatibility Complex class I - MHC I molecules. The MHC I presentation via is mostly associated with a cellular cytotoxic response and often do not elicit a satisfactory humoral response. One of the main strategies to target DNA-encoded antigens to the MHC II compartment is expressing the antigen within the Lysosome-Associated Membrane Protein (LAMP). The flavivirus envelope protein is recognized as the major virus surface protein and the main target for neutralizing antibodies. Different groups have demonstrated that co-expression of flavivirus membrane and envelope proteins in mammalian cells, fused with the carboxyl-terminal of LAMP, is able to induce satisfactory levels of neutralizing antibodies. Here we reviewed the use of the envelope flavivirus protein co-expression strategy as LAMP chimeras with the aim of developing DNA vaccines for dengue, West Nile and yellow fever viruses.

A vacinação é a estratégia mais prática e o melhor custo-benefício para prevenir a maioria das infecções dos flavivirus, para os quais existe vacina disponível. Entretanto, as vacinas baseadas em vírus atenuados podem potencialmente promover efeitos colaterais e, mais raramente, reações fatais. Diante deste cenário, o desenvolvimento de estratégias alternativas de vacinação, como vacinas baseadas em DNA codificando seqüências específicas dos flavivirus, está sendo considerado. Antí-genos citoplasmáticos endógenos, caracteristicamente codificados por vacinas de DNA plasmidial, são majoritariamente apresentados ao sistema imune através de moléculas do Complexo Maior de Histocompatibilidade de classe I - MHC I. A via de apresentação MHC I é mais associada à resposta celular citotóxica e, frequentemente, não elicita uma resposta humoral satisfatória. Uma das principais estratégias para direcionar antígenos codificados pelas vacinas de DNA para o compartimento MHC II é expressar estes antígenos dentro da Proteína de Associação à Membrana Lisossomal (LAMP). A proteína do envelope dos flavivirus é reconhecidamente a principal proteína de superfície viral e o principal alvo para anticorpos neutralizantes. Diferentes grupos têm demonstrado que a co-expressão das proteínas de membrana e do envelope dos flavivirus em células de mamíferos, fusionada com a porção carboxi-terminal de LAMP, é capaz de induzir níveis satisfatórios de anticorpos neutralizantes. Neste trabalho revisamos a estratégia de co-expressão da proteína do envelope dos flavivírus, como quimeras de LAMP, com o objetivo de desenvolver vacinas de DNA contra a febre do Oeste do Nilo, dengue e febre amarela.

DNA methylation profile in diffuse type gastric cancer: evidence for hypermethylation of the BRCA1 promoter region in early-onset gastric carcinogenesis

Diffuse type gastric carcinoma is the most aggressive type of gastric cancer. This type of tumor is not preceded by precancerous changes and is associated with early-onset and hereditary syndromes. To test the hypothesis that DNA methylation profile would be useful for molecular classification of the diffuse type gastric carcinoma, DNA methylation patterns of the CpG Island of 17 genes were studied in 104 cases and 47 normal adjacent gastric mucosa by Methylation-specific PCR, Immunohistochemistry and Hierarchicalclustering analysis. The most frequent methylated genes were FHIT, E-cadherin, BRCA1 and APC (>50%),followed by p14, p16, p15, p73, MGMT and SEMA3B (20-49%). Hierarchical clustering analysis reveals four groups with different clinical features. The first was characterized by hypermethylation of BRCA1 and younger age (<45 years old), and the second by hypermethylation of p14 and p16 genes, male predominance and Epstein-Barr virus infection. The third group was characterized by hypermethylation of FHIT and antrum located tumors and the fourth was not associated with any clinical variables. In normal adjacent mucosa only the p73 gene was significantly less methylated in comparison to tumor mucosa. DNA methylation identified subgroups of diffuse type gastric cancer. Hypermethylation of BRCA1 associated with young age suggests a role in early-onset gastric carcinoma.

Efectos de la infusión aguda de octreotido sobre la función renal en pacientes con cirrosis hepática e hipertensión portal / Effects of acute octreotide infusion on renal function in patients with cirrhosis and portal hypertension

Background: Octreotide is used in the treatment of acute variceal bleeding, based on its inhibitory effects of post-prandial splanchnic hyperemia and splanchnic venoconstriction. The consequences of these haemodynamic changes on renal circulation are not well known in cirrhotic patients. Aim: To evaluate the effects of acute octreotide administration on several parameters of renal function, including free water clearance, in patients with cirrhosis with or without ascites. Patients and Methods: Twenty cirrhotic patients, Child-Pugh A or B, with or without ascites, with esophageal varices, normal renal function and free of medications (vasoactive drugs or diuretics) were assigned to 2 different protocols. Protocol 1: 10 patients were randomized to receive octreotide or placebo, as a bolus followed by a continuous infusion. Glomerular filtration rate (GFR) and renal plasma flow (PRF) were measured, in basal conditions and during the drug or placebo administration. Protocol 2: 10 additional patients were randomized in the same way and free water clearance and urinary sodium excretion were again measured in the basal period and during the drug or placebo infusion. Results: After octreotide or placebo administration no significant changes were observed neither in GFR nor in PRF. The free water clearance decreased significantly during octreotide administration (3.12 ml/min±1.04 SE vs 0.88±0.39, p <.03). In both protocols no changes in mean arterial pressure were observed. Conclusions: Acute administration of octreotide to cirrhotic patients with portal hypertension, with or without ascites, did not produce any change in glomerular filtration rate or in estimated renal plasma blood flow. However the free water clearance decreased significantly. This effect, under chronic administration, could be clinically important and deserves further studies.

Helicobacter pylori: análisis de cagA y genotipificación de vacA en Chile: detección de una cepa s2/m1 / Helicobacter pylori: cagA status and vacA genotyping in Chile, detection of a s2/m1 strain

Background: The genes cagA and vacA encode H pylori virulence factors. Aim: To genotype these genes in H pylori strains isolated from patients with upper gastrointestinal symptoms. Material and methods: We studied 50 patients who underwent an upper gastrointestinal endoscopy, with positive culture for H pylori. Detection of cagA and vacA gerotyping was done using polymerase chain reactions. Results: The gene cagA was detected in 19 samples (38 per cent). Signal sequences s1 and s2 of vacA gene were detected in 16 samples each (32 per cent). There was simultaneous amplification of s1 and s2 in 6 samples and they were not detected in 9 samples. The middle region of vacA was m1 in 9 samples, m2 in 29 samples and there was simultaneous amplification of m1 and m2 in 12 samples. In 16 samples (32 per cent), more than one type of signal sequence or medial region was detected. Of those patients in whom vacA was the only genotype detected, 15 were s2/m2, 7 were s1/m1, 4 were s1/m2 and 1 was s2/m1. Conclusions: In these patients, the infection with cagA- H pylori strains, predominates, the prevalence of infection with s1 or s2 strains is similar and the predominant medial region is m2